LOCUS       sequence-272673-        7739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of C. elegans KNL1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7739)
  AUTHORS   Kern DM, Kim T, Rigney M, Hattersley N, Desai A, Cheeseman IM
  TITLE     The outer kinetochore protein KNL-1 contains a defined 
            oligomerization domain in nematodes.
  JOURNAL   Mol Biol Cell. 2015 Jan 15;26(2):229-37. doi: 
            10.1091/mbc.E14-06-1125. Epub 2014 Nov 19.
  PUBMED    25411336
REFERENCE   2  (bases 1 to 7739)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1091/mbc.E14-06-1125"; journalName: "Mol Biol Cell"; date: 
            "2015-01-15- 15"; volume: "26"; issue: "2"; pages: "229-37"
COMMENT     Sequence Label: pDK167
FEATURES             Location/Qualifiers
     source          1..7739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     35..54
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     138..157
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        138..156
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             230..252
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3299..3316
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3406..3424)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3420..3467
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3833..3838,3839..3855,3856..3861))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3888..3906)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3974..4078
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4079..4147,4148..4939)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4297..4316)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5110..5698
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5599..5618
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5852..5869
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5884..6023
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5949..5968)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6109..6131
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6125..6316)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
       61 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
      121 tctcgatccc gcgaaattaa tacgactcac tatagggaga ccacaacggt ttccctctag
      181 tgccggctcc ggagagctcc aattggaatt cgctagctct agaaataatt ttgtttaact
      241 ttaagaagga gatatacata tgtcgatgga gcctcgtaag aagcggaact cgattctcaa
      301 ggtgcggcaa gccgtcgaaa ccatcgagga aaccgtcatg aacagtgggc ctagttccac
      361 aacaactaat cgacgagtca gctttcataa cgtgaagcat gtcaagcagt atgacaggga
      421 ccatggtaaa attcttgacg ccacaccagt taaggagaag attactgaca ctattggatc
      481 agatggtatt ttgacaccac gtggcggaaa catggatatt tccgaatctc cggcctgcac
      541 gtcctcattt caagtgttcg gcggtggtaa tctcgataaa actatggata tgtctctcga
      601 aacaactatc aacgagaaca acgaaacggc gagattgttt gaaaccacaa gagatccaac
      661 agctgctgca gagaaagctg ccgaaaccac tactaaggct actgaacgtg ctgctagtat
      721 gccactggat gataccttag caatgttcaa tacaacgaat caagaagata aggatatgtc
      781 agttgatcgt tcagttcttt tcacgattcc caaagttccg aagcataacg ctacaatgaa
      841 tagaactata ccgatggacc tcgatgaatc aaaagcagcg ggcggccagt gcgatgaaac
      901 gatgaatgtg ttcaatttca caaacttgga agccgctgaa atggatacga gtaaattaga
      961 tgaaaataat accatgaatg ctatccggat tccgattaat tcaaacgtca tgcctgtaga
     1021 catggacatc actgaacatc acactttaat tgaagaaaag aaaaatgata cattcgggcc
     1081 aagtcaactg atggacattt cggcgccaca agttcaagtt aatgatactt tggccatttt
     1141 caacagtccg agagacatct gtaataaggg tttgggtgtt cctcagaatc taataaatat
     1201 cgcctcgaac gtcgtacctg tggacatgga catcactgat caggccgtat taaacgcgga
     1261 gaagaaaaat gatcaattcg agacaagtca gcttatggac atttctattc cgaaagttct
     1321 agtaaatgac actatggcga tgttcaacag cccgaaacac gtcagtaaga gcagcatgga
     1381 tctcgagaaa acgattgaag ccgctgacaa atcaacgaaa tacccgagta tcgcagatga
     1441 ggtggaagat ttagacatgg atatggatat cactgaacaa caaccatgtg aggctggtaa
     1501 tcagcagaac gacggcttgc aacttcaaaa ggaggattta atggacattt cggtgattcg
     1561 agattcacct gcagtaaacg acaccatggc tgtgttccag agtcctgcca gagtaaagat
     1621 cggagcgaac aactcgatca ttgattcgca gaaatctatc gtgttcggtg acgaaatgag
     1681 cattgacgag acacaaaatg atggaacctt gacgttgcca aagtcgaatg tagaagtgac
     1741 tacaactaat gatgtctaca cgtctctcga gcggcaagag gaaaatgctt cagaaaacgt
     1801 atccatgata aacgaatctt ctgttcattc ggaaatcgac aaaaagtcgt ttatgctcat
     1861 cgaagaagaa agggctttta tgcactcctc catgattgat gtagcacaaa agttggaaga
     1921 cgatggttcg tcgaagacgc cagtcatcct tgcttcacag tcagcttctc ttgccactaa
     1981 agaaccatca gcccttcaca actcgagtgc aactctcaac aattcgatgg aattggacaa
     2041 caatactctt cttaaaacta tgcaaattac aacgtgtgaa gacattagca tggtccatga
     2101 gtctattgct gttgaactga acagtaacaa agagcaggag caattcggag atgagacttt
     2161 gcagaaaaat gatacctcga atactggcgc gaatttcaca ttccaaggcc ataatgaaac
     2221 atcgcaaatc atgaacaatg tcgactcgga agcagtgaac acgtccaaga tttcaacata
     2281 ttcggctttc aatttgagca tcaaccagtc tatctctaaa cgacgtcgat ctcttctgaa
     2341 ttctgctcgt gaatctcctc gtcgtgttgc gttggagaat tctataatgt cgatgaatgg
     2401 gcaaacagtg gaagctctga cagaatatcg acagaataaa actatgcaga cgagtcaaga
     2461 ttcgatgccg agtatgagtt tgaacgattc gggaagagat attctcgcga tgaatacatc
     2521 agtccgctct cctcatctga attcttcaaa aactgctgcc ccaggaacac catcattgat
     2581 gtcacaaaat gtacaacttc cacccccatc tcctcaattc gaaatgccag acttcgatcc
     2641 agctgtggtc aacgttgtat atttaacatc tgaagatccg tccactgaac aacatccaga
     2701 agctctcaaa tttcagcgta ttgttgaaaa cgagaaaatg aaagtacaac acgagattga
     2761 ttctctgaat tcaaccaatc aactttctgc tgagaaaatt gatatgttga agactaagga
     2821 gctcttgaag tttagtcatg atgagcgaga agcgattatg attgcaagaa aagacgcgga
     2881 aatcaagttt ttggagcttc gtctgaaatt tgcactcgaa aaaaaaattg aaagtgacca
     2941 ggaaattgct gaactagaac aaggaaattc gaaaatggct gagcagctaa gaggtctcga
     3001 taagatggct gtcgttcaaa aagaactaga aaagctgaga agtcttcctc catcacgcga
     3061 agagagcggg aaaatccgaa aggagtggat ggagatgaag caatgggaat tcgaccagaa
     3121 aatgaaagca ctccgaaatg tacgctcaaa catgattgca cttcgttcag agaaaaatgc
     3181 tctcgaaatg aaagtcgcgg aagaacacga gaagtttgcc cagaggaacg atttgaagaa
     3241 aagtcgaatg ctggtgttct ctaaggctgt taagaaaatt gtgaacttcg gcagcagcca
     3301 tcatcatcat catcactagg gatccagatc taagcttggt accacgcgtg cgcgctgatc
     3361 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
     3421 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
     3481 ctatatccgg atatccacag gacgggtgtg gtcgccatga tcgcgtagtc gatagtggct
     3541 ccaagtagcg aagcgagcag gactgggcgg cggccaaagc ggtcggacag tgctccgaga
     3601 acgggtgcgc atagaaattg catcaacgca tatagcgcta gcagcacgcc atagtgactg
     3661 gcgatgctgt cggaatggac gatatcccgc aagaggcccg gcagtaccgg cataaccaag
     3721 cctatgccta cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat
     3781 ttcatacacg gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc
     3841 ttatcgatga taagctgtca aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg
     3901 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     3961 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4021 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4081 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4141 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4201 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4261 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4321 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4381 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4441 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4501 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4561 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4621 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4681 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4741 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4801 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4861 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4921 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4981 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5041 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5101 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5161 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5221 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     5281 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5341 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5401 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5461 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5521 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5581 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5641 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5701 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5761 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     5821 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     5881 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc
     5941 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     6001 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     6061 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     6121 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     6181 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     6241 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     6301 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga
     6361 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
     6421 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
     6481 tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
     6541 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt
     6601 ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg
     6661 ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag
     6721 taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc
     6781 gtggccagga cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg
     6841 cgatggatat gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat
     6901 tggctccaat tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt
     6961 cgaggtggcc cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc
     7021 ggcgcctaca atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt
     7081 gacgatcagc ggtccaatga tcgaagttag gctggtaaga gccgcgagcg atccttgaag
     7141 ctgtccctga tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc
     7201 gatgccgccg gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa
     7261 cgccagcaag acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc
     7321 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     7381 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     7441 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     7501 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     7561 tctcaagggc atcggtcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     7621 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
     7681 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctca
//
